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1.
Anal Bioanal Chem ; 412(24): 6407-6417, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32333077

RESUMO

Celiac disease (CD) is a chronic autoimmune disorder induced in genetically susceptible individuals by the ingestion of gluten from wheat, rye, barley, or certain varieties of oats. A careful diet follow-up is necessary to avoid health complications associated with long-term gluten intake by the celiac patients. Small peptides (GIP, gluten immunogenic peptides) derived from gluten digestion, which are excreted in the urine and feces, have emerged as promising biomarkers to monitor gluten intake. We have implemented a simple and sensitive label-free point-of-care (POC) device based on surface plasmon resonance for the direct detection of these biomarkers in urine. The assay employs specific monoclonal antibodies and has been optimized for the detection of the 33-mer α2-gliadin, known as the main immunogenic peptide of wheat gluten, and for the detection of GIP. Direct detection in undiluted urine has been accomplished by using biosensing chips containing a robust and stable biorecognition layer, obtained after carefully optimizing the biofunctionalization protocol. Excellent limits of detection have been reached (1.6-4.0 ng mL-1 using mAb G12 and A1, respectively), which ensures the detection of gluten peptides even when the gluten intake is around the maximum tolerable amount in the digestive tract (< 50 mg) for celiac individuals. No sample pretreatment, extraction, or dilution is required, and the analysis takes less than 15 min. The assays have excellent reproducibility' as demonstrated by measuring spiked urine samples containing the same target concentration using different biofunctionalized chips prepared and stored at different periods of time (i.e., CV% of 3.58% and 11.30%, for G12- and A1-based assays, respectively). The assay has been validated with real samples. These features pave the way towards an end-user easy-to-handle biosensor device for the rapid monitoring of gluten-free diet (GFD) and follow-up of the health status in celiac patients.


Assuntos
Doença Celíaca/urina , Dieta Livre de Glúten , Gliadina/urina , Fragmentos de Peptídeos/urina , Ressonância de Plasmônio de Superfície/instrumentação , Anticorpos Imobilizados/química , Anticorpos Monoclonais/química , Doença Celíaca/dietoterapia , Desenho de Equipamento , Humanos , Limite de Detecção , Ressonância de Plasmônio de Superfície/economia , Fatores de Tempo
2.
Biosens Bioelectron ; 119: 149-155, 2018 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-30125875

RESUMO

Acenocoumarol (Sintrom®) is an oral anticoagulant prescribed for the treatment of a variety of thromboembolic disorders such as atrial fibrillation and thrombosis or embolism. It inhibits fibrin production preventing clot formation. Acenocoumarol has a narrow therapeutic range, and its effects depend on several factors, such as body weight, age, metabolism, diet, certain medical conditions or the intake of additional drugs, among others. A higher dose may result in the risk of bleeding, while if it is too low, the risk of blood clot can increase. Complementary tools that allow the therapeutic drug monitoring (TDM) of acenocoumarol plasmatic levels from the starting of the treatment would be of paramount importance to personalize the treatment. Point-of-care (POC) devices can offer an added value in facilitating on-site monitoring (i.e. hospitals, primary care doctor or even by the patient itself) and can aid in dosage management. With this aim, we have developed a compact and simple nanoplasmonic sensing device based on gold nanodisks for the rapid monitoring of acenocoumarol, using highly specific polyclonal antibodies produced against this drug. A specific and reproducible label free indirect competitive assay has been developed and the viability of performing the evaluation directly in plasma diluted 1:1 has been demonstrated. A limit of detection (LOD) of only 0.77 ±â€¯0.69 nM, an IC50 of 48.2 ±â€¯5.12 nM and a dynamic range between 3.38 ±â€¯1.33 nM and 1154 ±â€¯437 nM were achieved, which easily fit within the drug plasma levels of acenocoumarol, making this approach a highly attractive option for its decentralized monitoring in human plasma.


Assuntos
Acenocumarol/sangue , Técnicas Biossensoriais/instrumentação , Análise Química do Sangue/instrumentação , Análise Química do Sangue/métodos , Monitoramento de Medicamentos/instrumentação , Monitoramento de Medicamentos/métodos , Anticoagulantes/sangue , Humanos , Limite de Detecção
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